Cell and enzyme compositions for modulating bile acids, cholesterol and triglycerides

Abstract

The invention relates to immobilized or encapsulated enzyme and / or cells to lower bile acids and cholesterol. The invention also relates to methods of quantitatively measuring bile acids. The invention provides a composition for decreasing the amount of a target compound in the gastrointestinal tract of an animal, comprising: a) a biologically active agent which decreases the amount of the target compound; b) a retainer for retaining the biologically active agent by contacting the agent to limit movement of the agent; and c) a carrier.

Description

FIELD OF THE INVENTION [0001] The invention relates to immobilized or encapsulated enzyme and / or cells to modulate bile acids, cholesterol and triglyceride levels in a subject. The invention also relates to methods of quantitatively measuring bile acids and triglycerides. BACKGROUND OF THE INVENTION [0002] Bile acids are important physiological agents that are required for the disposal of cholesterol and the absorption of dietary lipids and lipid soluble vitamins. Bile salts are the water-soluble end products of cholesterol, and are synthesized de novo in the liver. During normal enterohepatic circulation (EHC), the average bile salt pool is secreted into the duodenum twice during each meal, or an average of 6-8 times per day for the purpose of forming mixed micelles with the products of lipid digestion. During intestinal transit, most of the secreted bile salt is absorbed in the terminal ileum and is returned to the liver via the portal vein. The bile salt pool is replenished by he...

AI Technical Summary

Benefits of technology

[0012] ii) a retainer for retaining the biologically active agent, for example by immobilizing it on a surface and / or encapsulating it. This has the effect of isolating the agent and reducing its movement. The retainer optionally comprises a capsule, such as a capsule comprising a semi-permeable membrane, and / or a support, such as a polymer structure. The retainer is optionally a retainer means for retaining the agent; and

[0015] The invention also includes methods comprising contacting a biologically active agent (for example, a composition of the invention) with a target compound in an animal to, for example, degrade target compound in an animal to reduce target compound levels. The methods optionally modulate lipid or bile metabolism, with an agent such as BSH, for deconjugating bile acids to form target-degradation compounds. This has the effect of reducing bile acid levels. The methods optionally use a lipase, which breaks down lipids, such as triglycerides and their esters, to form target-degradation compounds such as fatty acids. The methods optionally use a cell, such as a bacterial cell, expressing the enzyme. The methods optionally involve oral administration or implantation in the animal. In the methods, the biologically active agent is optionally retained in a retainer, for example immobilized on a surface and / or encapsulated. This has the effect of isolating the agent and reducing its movement in the methods. The methods optionally further comprise collecting a target-degradation compound formed as a result of the agent's reaction with the target compound. In one embodiment, a bile acid is deconjugated and then, its by-product, DCA, is captured, for example by precipitation and collection in a capsule, where it is held until it is excreted.

[0016] In one embodiment of the invention, the present inventors have shown that immobilized or encapsulated genetically engineered cells, such as Lactobacillus plantarum 80 cells expressing BSH, are a biologically active agent that efficiently hydrolyzes bile acids and that are useful in the deconjugation of human bile acids.

[0023] Naturally occurring bacteria can significantly lower serum cholesterol levels by hydrolyzing bile salts in the intestinal tract. Accordingly, the present invention provides a method for lowering of serum cholesterol of patients, which comprises administering a bile acid lowering amount of a composition of the present invention.

[0039] In one embodiment, the retainer limits agent movement by a retainer surface immobilizing the agent and / or by the retainer encapsulating the agent. In a further embodiment, the retainer encapsulates the agent and reduces exposure of the biologically active agent to antibodies and permits exposure of the biologically active agent to nutrients. The retainer optionally comprises a semi-permeable membrane. The semi-permeable membrane also optionally comprises a MWCO of about 3000 D to 950,000 D In one embodiment, the retainer comprises a polymer bead and the agent is immobilized on the bead.

Problems solved by technology

Though effective, these methods still have several limitations.

Further, oral administration of live bacterial cells can pose problems.

In addition, risk of systemic infections, deleterious metabolic activities, adjuvant side-effects, immuno-modulation and risk of gene transfer has limited their use (De Boever and Verstraete, 1999; Christiaens et al., 1992).

Metabolic activities and immuno-modulation, have limited its clinical use (De Boever et al., 2000).

As well, when ileal transport of bile acids is defective due to a congenital defect, resection of the ileum, or disease, elevated intraluminal concentrations of bile acids can induce the secretion of electrolytes and water causing diarrhea and dehydration.

These methods have several limitations.

Such therapies are often sub-optimal and carry a risk for serious side effects.

However, the combined restricted vegetarian diet (free of meat, fish, chicken, vegetable oils and all dairy fat products) and aerobic approach, is unrealistic for all but the most dedicated individuals.

In addition, none of these dietary interventions have been shown to arrest or cure atherosclerosis or other high blood cholesterol associated diseases.

Although effective at reducing cholesterol levels, they are nevertheless expensive (Attanasio et al., 2001; Hodgson and Cohen, 1999; Prosser et al.

The therapeutic potential of free bacteria is hampered by inherent limitations in their use.

Also, oral administration of live bacterial cells can cause a host immune response, and can be detrimentally retained in the intestine replacing the natural intestinal flora (Taranto et al., 2000; Chin et al., 2000; De Boever and Verstraete, 1999).

Furthermore, there are some practical concerns regarding the production, cost, and storage of products containing free bacteria (De Boever and Verstraete, 1999).

Thus, concerns of safety and practicality have prevented the regular use of this promising therapy in clinical practice.

Other problematic diseases or disorders arise from disrupted lipid metabolism.

The resulting condition, known as exocrine or pancreatic insufficiency, leads to weight loss and very foul-smelling stools or diarrhea.

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